BioLife Solutions Inc is a company specializing in Biological Packaging and dedicated to developing new technologies for hypothermic storage (4 to 8C) and cryopreservation (-196C) of human cells and tissues. The HypoThermosol (HTS) platform consists of HTS-PURGE, HTS-BASE, HTS-DCC, HTS-FRS and other custom preservation solutions being utilized by clients in the Regenerative Medicine market. BioLife's serum-free CryoStor cryopreservation solution platform is designed for cell and tissue storage in liquid nitrogen and requires reduced DMSO levels. BioLife's solutions are designed around an understanding of the molecular events underlying cell death cascades that can be launched due to preservation storage. Once known, solution formulation is altered in such a manner that inhibits these cell death processes. Several of BioLife's HTS variants have been shown to have a marked benefit over present commercial solutions (e.g.ViaSpan) designed for organ storage. BioLife's CryoStor series may be better at cryopreservation than the current ATCC industry protocol. BioLife's solutions are now a part of BioHeart's FDA-approved IND in cellular cardiomyoplasty - a successful international program in cardiac Regenerative Medicine made possible, in part, by BioLife's HTS-FRS. BioLife has recognized that OptisoI-GS, a corneal storage solution distributed by Chiron Vision, was developed 12 years ago and its formulation is not based on the molecular events that occur to corneal cells subjected to preservation. Under this SBIR program, BioLife will develop specialized corneal preservation technologies, HTS-CORNEA and CryoStor-CORNEA, that will provide enhanced performance over that of OptisoI-GS. The Phase I project proposes to: (1) Develop a cell culture method and multiple endpoint assay system to assess bovine and human corneal cells in vitro and their responses to HTS and CryoStor variants; (2) Determine the relative contributions of apoptosis and necrosis in preservation-induced, corneal cell death; (3) Determine if the addition of chondroitin sulfate, an extracellular matrix component molecule present in OptisoI-GS and other corneal storage solutions, is necessary for HTS-CORNEA and CryoStor-CORNEA to maintain cell and tissue protection in the cold; and (4) Test the inclusion of MMP and NO inhibitors as additives to HTS-CORNEA and CryoStor-CORNEA. The Phase II project will test HTS-Cornea and CryoStor-Cornea on the preservation of whole, human corneas. [unreadable] [unreadable] [unreadable]